Fig 3.

VPS4B depletion prolongs EGFR expression and signaling in EGF-treated SKBR3 cells. (A) Parental and VPS4B-knockdown (sh-VPS4B) SKBR3 cells in 2D (left) and 3D (right) cultures were serum starved for 16 h. Cells were subsequently treated with EGF (100 ng/ml) for the indicted times prior to harvesting. Equal amounts of whole-cell lysates were subjected to Western blot analyses with the indicated antibodies. (B) SKBR3 and SKBR3/sh-VPS4B cells were treated with EGF as for panel A and immunostained with primary antibody and the corresponding fluorescent secondary antibodies to detect EGFR (green) and Rab7 (red). Bar, 10 μm.